SpringerLink
Log in

Screening and Constructing of Novel Angiotensin I-Converting Enzyme Inhibiting Peptides from Walnut Protein Isolate and Their Mechanisms of Action: A Merged In Silico and In Vitro Study

  • Research
  • Published:
Plant Foods for Human Nutrition Aims and scope Submit manuscript

Abstract

Angiotensin I-converting enzyme (ACE)-inhibiting peptides were isolated from walnut protein isolate (WPI) using ultrasound-assisted extraction. This study aimed to assess the impact of ultrasonic pretreatment on the physicochemical properties of WPI. The optimal extraction conditions for WPI were determined as a 15-min ultrasonic treatment at 400 W. Subsequently, the hydrolysate exhibiting the highest in vitro ACE-inhibiting activity underwent further processing and separation steps, including ultrafiltration, ion exchange chromatography, liquid chromatography-tandem mass spectrometry, ADMET screening, and molecular docking. As a result of this comprehensive process, two previously unidentified ACE-inhibiting peptides, namely Tyr-Ile-Gln (YIQ) and Ile-Tyr-Gln (IYQ), were identified. In addition, a novel peptide, Ile-Lys-Gln (IKQ), was synthesized, demonstrating superior ACE-inhibiting activity and temperature stability. In silico analysis estimated an in vivo utilization rate of 21.7% for IKQ. These peptides were observed to inhibit ACE through an anti-competitive mechanism, with molecular docking simulations suggesting an interaction mechanism involving hydrogen bonding. Notably, both IYQ and IKQ peptides exhibited no discernible toxicity to HUVECs cells and promoted nitric oxide (NO) generation. These findings underscore the potential of ultrasonicated WPI in the separation of ACE-inhibiting peptides and their utility in the development of novel ACE inhibitors for functional food applications.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Subscribe and save

Springer+ Basic
EUR 32.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or Ebook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Price includes VAT (United Kingdom)

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

Data Availability

The data from the current study can be obtained from the corresponding author upon reasonable request.

References

  1. Mills KT, Stefanescu A, He J (2020) The global epidemiology of hypertension. Nat Rev Nephrol 16:223–237. https://doi.org/10.1038/s41581-019-0244-2

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Li Z, He Y, He H et al (2023) Purification identification and function analysis of ACE inhibitory peptide from Ulva prolifera protein. Food Chem 401:134127. https://doi.org/10.1016/j.foodchem.2022.134127

    Article  CAS  PubMed  Google Scholar 

  3. Daskaya-Dikmen C, Yucetepe A, Karbancioglu-Guler F et al (2017) Angiotensin-I-converting enzyme (ACE)-inhibitory peptides from plants. Nutrients (9). https://doi.org/10.3390/nu904031i

  4. ** F, Wang Y, Tang H et al (2020) Limited hydrolysis of dehulled walnut (Juglans regia L.) proteins using trypsin: functional properties and structural characteristics. LWT - Food Sci Technol 133:110035. https://doi.org/10.1016/j.lwt.2020.110035

  5. Tang HK, Wang C, Cao SN et al (2022) Novel angiotensin I-converting enzyme (ACE) inhibitory peptides from walnut protein isolate: separation, identification and molecular docking study. J Food Biochem 46(12):e14411. https://doi.org/10.1111/jfbc.14411

    Article  CAS  PubMed  Google Scholar 

  6. Liu DD, Guo YT, Ma HL (2022) Production, bioactivities and bioavailability of bioactive peptides derived from walnut origin by-products: a review. Crit Rev Food Sci Nutr 1:1–16. https://doi.org/10.1080/10408398.2022.2054933

    Article  CAS  Google Scholar 

  7. Feng L, Peng F, Wang X et al (2019) Identification and characterization of antioxidative peptides derived from simulated in vitro gastrointestinal digestion of walnut meal proteins. Food Res Int 116:518–526. https://doi.org/10.1016/j.foodres.2018.08.068

    Article  CAS  PubMed  Google Scholar 

  8. Gu M, Chen HP, Zhao MM et al (2015) Identification of antioxidant peptides released from defatted walnut (Juglans sigillata dode) meal proteins with pancreatin. LWT - Food Sci Technol 60(1):213–220. https://doi.org/10.1016/j.lwt.2014.07.052

  9. Fang SX, Ruan GR, Hao J et al (2020) Characterization and antioxidant properties of manchurian walnut meal hydrolysates after calcium chelation. LWT - Food Sci Technol 130:109632. https://doi.org/10.1016/j.lwt.2020.109632

    Article  CAS  Google Scholar 

  10. Yang X, Wang L, Zhang F et al (2020) Effects of multi-mode S-type ultrasound pretreatment on the preparation of ACE inhibitory peptide from rice protein. Food Chem 331:127216. https://doi.org/10.1016/j.foodchem.2020.127216

    Article  CAS  PubMed  Google Scholar 

  11. Zaharuddin ND, Barkia I, Wan Ibadullah WZ et al (2022) Identification, molecular docking, and kinetic studies of six novel angiotensin-I-converting enzyme (ACE) inhibitory peptides derived from Kenaf (Hibiscus cannabinus L.) seed. Int J Biol Macromol 220:1512–1522. https://doi.org/10.1016/j.ijbiomac.2022.09.142

  12. Majumder K, Wu J (2010) A new approach for identification of novel antihypertensive peptides from egg proteins by QSAR and bioinformatics. Food Res Int 43:1371–1378. https://doi.org/10.1016/j.foodres.2010.04.027

    Article  CAS  Google Scholar 

  13. Mirzapour M, Rezaei K, Sentandreu MA (2017) Identification of potent ACE inhibitory peptides from wild almond proteins. J Food Sci 82:2421–2431. https://doi.org/10.1111/1750-3841.13840

    Article  CAS  PubMed  Google Scholar 

  14. Huang Y, Jia F, Zhao J et al (2021) Novel ACE inhibitory peptides derived from yeast hydrolysates: Screening, inhibition mechanisms and effects on HUVECs. J Agric Food Chem 69(8):2412–2421. https://doi.org/10.1021/acs.jafc.0c06053

    Article  CAS  PubMed  Google Scholar 

  15. Chen MT, Wang L, Zheng C, Ma A et al (2023) Novel ACE inhibitory peptides derived from bighead carp (Aristichthys nobilis) hydrolysates: screening, inhibition mechanisms and the bioconjugation effect with graphene oxide. Food Biosci 52:102399. https://doi.org/10.1016/j.fbio.2023.102399

  16. Lee JH, Kim TK, Yong HI et al (2023) Peptides inhibiting angiotensin-I-converting enzyme: isolation from flavourzyme hydrolysate of Protaetia brevitarsis larva protein and identification. Food Chem 399:133897. https://doi.org/10.1016/j.foodchem.2022.133897

    Article  CAS  PubMed  Google Scholar 

  17. Helmlinger J, Sengstock C, Groß-Heitfeld C et al (2016) Silver nanoparticles with different size and shape: equal cytotoxicity, but different antibacterial effects. RSC Adv 6:18490–18501. https://doi.org/10.1039/C5RA27836H

    Article  CAS  ADS  Google Scholar 

  18. Sangsawad P, Roytrakul S, Yongsawatdigul J (2017) Angiotensin converting enzyme (ACE) inhibitory peptides derived from the simulated in vitro gastrointestinal digestion of cooked chicken breast. J Funct Foods 29:77–83. https://doi.org/10.1016/j.jff.2016.12.005

    Article  CAS  Google Scholar 

  19. Crespo MI, Chabán MF, Lanza PA et al (2019) Inhibitory effects of compounds isolated from Lepechinia meyenii on tyrosinase. Food Chem Toxicol 125:383–391. https://doi.org/10.1016/j.fct.2019.01.019

    Article  CAS  PubMed  Google Scholar 

  20. Choi JH, Lee GH, ** SW et al (2021) Impressic acid ameliorates atopic dermatitis-like skin lesions by inhibiting ERK1/2-mediated phosphorylation of NF-κB and STAT1. Int J Mol Sci 22:2334. https://doi.org/10.3390/ijms22052334

  21. Gülseren İ, Güzey D, Bruce BD et al (2007) Structural and functional changes in ultrasonicated bovine serum albumin solutions. Ultrason Sonochem 14:173–183. https://doi.org/10.1016/j.ultsonch.2005.07.006

    Article  CAS  PubMed  Google Scholar 

  22. Lee SY, Hur SJ (2017) Antihypertensive peptides from animal products, marine organisms, and plants. Food Chem 228:506–517. https://doi.org/10.1016/j.foodchem.2017.02.039

    Article  CAS  PubMed  Google Scholar 

  23. Ferreira LG, Andricopulo AD (2019) ADMET modeling approaches in drug discovery. Drug Discov Today 24:1157–1165. https://doi.org/10.1016/j.drudis.2019.03.015

    Article  CAS  PubMed  Google Scholar 

  24. Qian B, Tian C, Huo J et al (2019) Design and evaluation of four novel tripeptides as potent angiotensin converting enzyme (ACE) inhibitors with anti-hypertension activity. Peptides 122:170171. https://doi.org/10.1016/j.peptides.2019.170171

    Article  CAS  PubMed  Google Scholar 

  25. Sharma S, Pradhan R, Manickavasagan A et al (2022) Exploration of corn distillers solubles from selective milling technology as a novel source of plant-based ACE inhibitory protein hydrolysates. Food Chem 388:133036. https://doi.org/10.1016/j.foodchem.2022.133036

    Article  CAS  PubMed  Google Scholar 

  26. Han R, Maycock J, Murray BS et al (2019) Identification of angiotensin converting enzyme and dipeptidyl peptidase-IV inhibitory peptides derived from oilseed proteins using two integrated bioinformatic approaches. Food Res Int 115:283–291. https://doi.org/10.1016/j.foodres.2018.12.015

    Article  CAS  PubMed  Google Scholar 

  27. Li R, Zhou X, Sun L et al (2022) Identification, in silico screening, and molecular docking of novel ACE inhibitory peptides isolated from the edible symbiot boletus griseus-hypomyces chrysospermus. LWT - Food Sci Technol 169:114008. https://doi.org/10.1016/j.lwt.2022.114008

    Article  CAS  Google Scholar 

  28. Pinciroli M, Aphalo P, Nardo AE et al (2019) Broken rice as a potential functional ingredient with inhibitory activity of renin and angiotensin-converting-enzyme (ACE). Plant Foods Hum Nutr 74:405–413. https://doi.org/10.1007/s11130-019-00754-6

    Article  CAS  PubMed  Google Scholar 

  29. Gilmore AR, Alderdice M, Savage KI et al (2019) ACE: A workbench using evolutionary genetic algorithms for analyzing association in TCGA. Cancer Res 79:2072–2075. https://doi.org/10.1158/0008-5472.CAN-18-1976

    Article  CAS  PubMed  Google Scholar 

  30. Fan Y, Yu Z, Zhao W et al (2020) Identification and molecular mechanism of angiotensin-converting enzyme inhibitory peptides from larimichthys croceatitin. Food Sci Hum Well 9:257–263. https://doi.org/10.1016/j.fshw.2020.04.001

    Article  Google Scholar 

  31. Liang F, Shi Y, Shi J et al (2021) A novel angiotensin-I-converting enzyme (ACE) inhibitory peptide IAF (ile-ala-phe) from pumpkin seed proteins: in silico screening, inhibitory activity, and molecular mechanisms. Eur Food Res Technol 247:2227–2237. https://doi.org/10.1007/s00217-021-03783-1

  32. Li J, Huo X, Zheng Y et al (2023) ACE-inhibitory peptides identified from quinoa bran Glutelin-2 hydrolysates. In silico screening and characterization, inhibition mechanisms of ACE, coordination with zinc ions, and stability. Plant Foods Hum Nutr 78:419–425. https://doi.org/10.1007/s11130-023-01074-6

  33. Liu C, Fang L, Min W et al (2018) Exploration of the molecular interactions between angiotensin-I-converting enzyme (ACE) and the inhibitory peptides derived from hazelnut (Corylus heterophylla Fisch.). Food Chem 245:471–480. https://doi.org/10.1016/j.foodchem.2017.10.095

  34. Forghani B, Zarei M, Ebrahimpour A et al (2016) Purification and characterization of angiotensin converting enzyme-inhibitory peptides derived from Stichopus horrens: stability study against the ACE and inhibition kinetics. J Funct Foods 20:276–290. https://doi.org/10.1016/j.jff.2015.10.025

    Article  CAS  Google Scholar 

  35. Pereira AM, Lisboa CR, Santos TD et al (2019) Bioactive stability of microalgal protein hydrolysates under food processing and storage conditions. J Food Sci Technol 56:4543–4551. https://doi.org/10.1007/s13197-019-03915-2

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Funding

This study was supported by a Major Science and Technology Project of **njiang Uygur Autonomous Region (2022A02009-4) and a National Key Research and Development Project (2022YFD1000104-03).

Author information

Author notes

  1. Yuzhen Wang, Hengkuan Tang these authors contributed equally to this work.

Authors and Affiliations

  1. State Key Laboratory of Efficient Production of Forest Resources, Bei**g Key Laboratory of Forest Processing and Safety, College of Biological Sciences and Technology, Bei**g Forestry University, No. 35, Tsinghua East Road, Haidian District, Bei**g City, 100083, China

    Yuzhen Wang, Hengkuan Tang, **nyue Deng, Yijie Shen, Mingjian Tang & Fengjun Wang

  2. The Institute of Inspection and Supervision, Hygiene and Health in Chaoyang District of Bei**g, Bei**g, 100021, China

    Hengkuan Tang

Authors
  1. Yuzhen Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Hengkuan Tang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. **nyue Deng
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Yijie Shen
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Mingjian Tang
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Fengjun Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

Yuzhen Wang: Conceptualization, Experiments, Methodology, Formal analysis, Writing—original draft, Investigation. Hengkuan Tang: Experiments, Formal analysis, Software. **nyue Deng: Data curation, Formal analysis. Yijie Shen: Data curation, Formal analysis. Mingjian Tang: Software, Data curation. Fengjun Wang: Conceptualization, Funding acquisition, Resources, Supervision, Validation.

Corresponding author

Correspondence to Fengjun Wang.

Ethics declarations

Competing interests

The authors declare no competing interests.

Ethics Approval

This article does not include any studies with human or animals’ subjects.

Consent to Participate

Not applicable.

Consent for Publication

Not applicable.

Conflict of Interest

The authors declare no conflict of interest.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary Information

Below is the link to the electronic supplementary material.

Supplementary file1 (DOCX 432 KB)

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Wang, Y., Tang, H., Deng, X. et al. Screening and Constructing of Novel Angiotensin I-Converting Enzyme Inhibiting Peptides from Walnut Protein Isolate and Their Mechanisms of Action: A Merged In Silico and In Vitro Study. Plant Foods Hum Nutr 79, 48–58 (2024). https://doi.org/10.1007/s11130-023-01122-1

Download citation

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11130-023-01122-1

Keywords

Search

Navigation